乙酸异戊酯高产菌株脂肪酶基因Lip2的克隆及表达研究

Study on the cloning and prokaryotic soluble expression of Lipase gene Lip2 of high- yield isoamyl acetate strain

摘要: 通过PCR扩增了乙酸异戊酯高产菌Loq-c的脂肪酶基因Lip2,其序列与Gene Bank中的Candida cylindracea Lip2 gene（序列号X64704.1）的序列相似度最高,为89%。构建原核表达载体p QE-lip2,转化大肠杆菌E.coli BL21（DE3）,获得了重组菌BL21（p QE-lip2）。经过表达条件探索,用SDS聚丙烯酰胺凝胶电泳分析,脂肪酶Lip2原核可溶表达最佳条件为:诱导温度30℃,IPTG使用浓度为1.5 mmol/L,150 r/min转速的条件下诱导表达5 h。

Abstract: The gene encoding Lipase2( Lip2) from high- yield isoamyl acetate strain Loq- c was amplified by polymerase chain reaction( PCR),its sequence was most similar to Candida cylindracea Lip2 gene( X64704.1),the similarity was 89%.Subsequently,the corresponding expression vector p QE- lip2 was constructed and cloned into E.coli BL21( DE3),resulting in recombinant strain BL21( p QE- lip2).After explorations,SDS- PAGE analysis showed that the best conditions for soluble expression of Lip2 was: 30 ℃,IPTG 1.5 mmol / L,150 r / min,induced for 5 hours.